In vivo method for producing female offsprings in bovines

ABSTRACT

The invention provides an in&#39;vivo method whereby female offsprings can be produced in bovines said method comprising the step of administering a therapeutically effective amount of a material comprising acetic acid, or its pharmaceutically acceptable derivatives to female bovines just after or before insemination.

This application is a National Stage application filed under 35 U.S.C371 of International application PCT/IN00/00125, filed Dec. 15, 2000,which designates the U.S. ,the entire contents of which are herebyincorporated by reference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to an in vivo method for producing femaleoffsprings in mammals. More specifically, it relates to a novel methodfor alteration of sex ratio to produce female offsprings, especially incase of bovine and such other animals.

BACKGROUND AND PRIOR ART OF THE INVENTION

Sex determination has been a favorite subject for reproductivephysiologists since long. Sex determination is a process whereby the sexof the offspring is decided by sex chromosomes in mammals and by factorssuch as temperature in certain chordates such as reptiles. Sexassignment and determination in mammals is a method whereby the sex ofthe offspring is decided even before actual formation of the zygote bythe fusion of an ovum with a spermatozoa bearing the sex chromosome of aparticular sex and conditions are created such that the combination ofchromosomes leading to the formation of a fetus of desired sex. Sexratio is an indicator/measure devised to ascertain the proportion ofmales and females in a given population. There have been severalattempts in the prior art to alter the sex ratio in mammals.

Some chordate species are known to be temperature dependent for sexualdetermination (TSD) (Bull, J. J 1980, Qrtly Rev Biol, 53, 3-21, 1980).In case of such animals, the sex of the offspring instead of beingdetermined by sex chromosomes, is determined by the temperature at whichegg is incubated. Further, there are certain species where applicationof certain hormones alters the sex of the offspring. Such speciesinclude certain reptiles and ratites. Such attempts to alter the sex arediscussed and disclosed in U.S. Pat. Nos. 5,201,280 and 5,377,618.

However, in case of mammals, sex of the offspring is determined byrandom combinations of X- or Y-chromosome bearing sperms with an ovumalways containing the X-chromosome and giving rise to a sex ratio ofalmost 50:50. In case of mammals also, some workers have tried tomanipulate sex ratios. Manipulation of sex ratio gains prominence withthe fact that mammals are an important family and success in alteringthe sex ratio in mammals, especially to the female side, has advantagesin milk and meat producing species and in evolving livestock of betterquality. This becomes doubly important in cross-breeding programs where50% of the offsprings turn out to be of female sex and remaining 50% ofmale sex. The offsprings of the male sex are not generally favoured inthe livestock industry. Altering the sex ratio, thereby leading topreferential production of females may make such cross-breeding programsa tremendous success. This also keeps the proportion of male populationsof these species to the minimum extent as required for inseminationpurposes only.

In the light of these advantages, many workers have attempted to altersex ratios throughout the last century. A number of attempts made about1940 were based on the assumption that vaginal pH controlled the sex ofthe offspring. Schroeder V. (Physico-chemical methods of sex regulationof the progeny of mammals. Abstr in J. Hered 1941: 32:248) discussedefficacy of physico-chemical methods to regulate sex ratios. Butconclusive evidence of efficacy of these methods was not found incareful scientific investigations (Salisbury and VanDemark: Physiologyof reproduction and artificial insemination in cattle, 1961).

Later on, this approach of change in vaginal pH was deserted andreproductive physiologists concentrated on attempts to produce sexedsemen. A sexed semen contains either X- or Y- bearing sperms in completeor accentuated concentrations, and which, when combined with ova,containing X-chromosome, either female (XX) or male (XY) offsprings areproduced in complete or relatively greater proportions.

Several workers have carried on research on this subject and useddifferent techniques to separate X- and Y- bearing sperms. (The spermsonly contain X-Y-chromosomes, however, the applicant has referred thesperms as X- or Y- for convenience.) Lindahl in 1953 reported success inproducing sexed semen in bulls (Counter-streaming of bull spermatozoa,Nature: 1956, 178: 491-92). Gordon M. J. (Scientific American, 199:87-94, 1958) also discussed a method to control the sex ratio. Thisstudy represented yet another approach to produce sexed semen.Bhattacharaya et al (An Attempt to determine the sex ratio of calves byartificial insemination with spermatozoa separated by sedimentation,Nature 211: 863: 1966), were able to achieve a degree of success inproducing sexed semen in bulls. Ericsson (Isolation of fractions rich inhuman Y-sperm, Nature 1973: 246:421) reported a method to get fractionsricher in human Y-sperms. Gledhill (Control of mammalian sex by sexingthe sperm: Fertil. Steril. 1983, 40(5): 572-74) and Corson et al (“Sexselection by sperm separation and insemination”, Fertil. Steril, 1984:42:756) also reported new methods to produce sexed semen. However, nopredictable and repeatable methodology could be evolved by these workersresulting in significant shift in sex ratio (Hunter, Reproduction ofFarm Animals, 1982, 138-139), has also stated that despite all theseattempts, modification of sex ratio still remains a mirage on thehorizon. The reason behind this may be that sperms are haploid cells andhaploid cells as distinct from diploid cells express a change in geneticconstitution in surface characteristics, is still not clear (Hunter,1982). Also, as pointed by Hafez (Reproduction in Farm Animals, fifthedition, 1987, 499), these attempts were hampered by the lack oflaboratory tests to evaluate the degree of sperm separation.

Sex determination was reported also on a different line. Certaincompounds, hormones, sera etc. were reported to have an effect on thealtering of sex ratio. Bennett & Boyce, (Sex ratio in progeny of miceinseminated with sperm treated with HY antiserum, Nature, 246:308, 1973)reported that insemination with sperm treated with antisera to aY-linked histocompatibility antigen produced 45.4% males compared with53.4% for controls in mice. Barrat and Leger (J. Gyneiol. Obstet. Biol.Reprod, Paris, 8, 332, 1979) reported that administration of clomiphenecitrate and/or gonadotropins resulted in 8.7% lowering of sex ratio.Beernik et al (“Factors influencing human sex ratio,” presented at theAnnual meeting of American Fertility Society, 1984) reported similarresults for humans. Sampson et al (“Gender After induction of Ovulationand artificial insemination” : Fertil. Steril. 40; pg 481, 1983)reported that with the induction of super ovulation, multiple birthsshowed a marked skewness towards male births. Mitra & Chowdhary (Abstrin animal Breed, 58(4): No.2354, 1990) showed that glyceryl phosphoruscholine diesterase activity of uterine fluid had an effect in alteringthe secondary sex ratio (i.e. at birth) in rats.

Thus, the prior art is replete with attempts to control the sex ofmammalian offspring because the outcome of this research is valuable ina variety of economic conditions, for example, preferential birth offemale calves in dairy herds would improve the rate of achievingsuperior animal strains by selective breeding. It would also be valuablemedically where for example it is desired to prevent the birth of sonsto mothers who are carriers of a genetic diseases which affect onlymales.

Each of the above attempts in the prior art represent differentapproaches towards sex assignment or production of off-springs of adesired sex. Recognizing the need to develop a simple and easy methodfor the assignment of sex or production of offsprings of significantnumber of females, the applicants conducted a thorough investigation onvarious chemicals capable of sex assignment and determination inoffsprings. With the singular objective of increasing female populationin livestock, the applicants screened a few chemicals and to theirsurprise found that a material essentially containing an acetyl group,such as vinegar, could be successfully used to achieve the abovepurpose. In addition, the applicant has arrived at a methodology toobtain female population, which is very cost-effective, easy to performand does not involve in vitro treatment of sperms.

OBJECTS OF THE INVENTION

The main objective of the invention is to provide an in vivo methodwhereby significant female offsprings can be produced in mammals.

Another objective is to provide a method whereby the female populationof livestock can be increased, to make cross-breeding programs withlivestock of exotic breeds a tremendous success.

SUMMARY OF THE INVENTION

Accordingly, the invention provides an in vivo method whereby femaleoffsprings can be produced in mammals, especially the bovine species,said method comprising the step of administering a therapeuticallyeffective amount of dilute acetic acid, or its pharmaceuticallyacceptable derivatives to female mammals just after insemination.

DETAILED DESCRIPTION OF THE INVENTION

The present invention in its broadest aspect relates to a method forpreferential production of female offsprings in mammals. The method ofthe invention is specifically applicable to members of the bovine familysuch as cows and buffaloes and other animals such as horses, sheep,dogs, goats, etc. It is the Applicant's finding that administration ofdilute acetic acid, or its derivatives, (hereinafter as “the material”for sake of brevity) to a mammal, within about 30 minutes afterinsemination will lead to preferential production of female offsprings.In fact, it is the Applicant's experience that the offspring producedafter such administration is generally female.

The method of the invention comprises the steps of insemination,artificial or natural of the female animal and administration of atherapeutically effective amount of the material, comprising essentiallyof a combination of acetic acid, its derivatives to the animalimmediately after insemination. Thereafter, the animal is allowed to eatand resume its usual activities in its natural surroundings andenvironment.

By “therapeutically effective amount”, the applicants imply an amountthat will enable production of female offsprings. Again, the dosage orthe amount of material to be administered will vary from animal toanimal and can be readily determined by a person skilled in the art onthe basis of body weight of the subject to which the material is to beadministered. For the instance, in case of cows and buffaloes, theamount may be about 150 to 800 ml of the said material. Preferably, 250to 400 ml of the material may be administered to the animal. In short,the amount administered should be such that it is not lethal to theanimal.

It is the Applicant's finding that the material that causes productionof female offsprings in mammals is a substance essentially comprisingacetyl radical. Typical examples of such material are vinegar, diluteacetic acid, sodium acetate and the natural or synthetic derivativesthereof. The material may be obtained from natural sources or derived bysynthetic methods.

The preferred material is vinegar. The essence of the invention lies inthe use of a material rich in or essentially comprising acetic acid foradministration to a mammal. The material known to be rich in acetic acidor vinegar as readily available. While the use of other such materialfalls within the scope of the invention. the applicants recommend theuse of vinegar prepared by prolonged fermentation. The natural sourcesof vinegar are crushed beet juice, sugar cane juice, molasses etc. Suchjuices are subjected to extended periods of fermentation such as 2-8months depending upon the season of the year. The fermentation may bepreferably carried out in any earthenware under optimum environments. Ina preferred embodiment, fermenting agents such as vinegar madepreviously by the same process or any other such fermenting agent may beadded to the broth. However, with the application of modem technology,this preparation of vinegar can be effected within a short period oftime, i.e. within 10 to 20 days or so, depending upon the quality offermenting agents and physical and chemical environments maintained. Thematerial may be decanted at regular intervals to avoid contamination andgrowth of unwanted organisms. The material produced at the end of such aprocess is essentially rich in acetic acid, and also contains traces ofacetaldehyde, acetic anhydride and ethanol.

The materials that can be used for administration to the animal includedilute acetic acid, sodium or potassium acetate in acidic pH, bothsolutions preferably kept at a pH of about 3, the natural or syntheticderivatives thereof.

The insemination of the mammal may be effected naturally or by adoptionof artificial methods as known in the art. In an embodiment, thismaterial is administered to the animal as early as possible afterinsemination. The step of administration of the material to the animalmay even precede the process of insemination as an alternate embodiment.In case of animals where repeated insemination occurs, such as in dogs,care is taken to ensure that the insemination is restricted to onceonly. It must also be ensured that the insemination is not subjected toprolonged or extended periods of time so that the peak levels formaximum effects of the material during the process of fertilization arerealized.

The time period for the administration of the material to the animal isquite critical. The period may of course vary from one animal to theother, but the general thumb rule is administration of the materialwithin 30 minutes after insemination. The reason is that the materialmust be administered to the animal before zygote formation. In any case,the material should be administered at least within one hour afterinsemination. If the material is to be administered before insemination,then it may be administered 1 or 3 hours before insemination.

After administering the material as a single dose, the fetus developsnormally and the animal goes through, completes pregnancy and givesbirth to totally normal and viable female offspring/s. It is found thatthe offsprings produced according to the method of the invention lead anormal life. Also these offsprings when mated with normal males, produceviable offsprings. The applicants have observed that administration ofthe material to the animal does not evoke any adverse reactions orside-effects like fever, skin reactions, behavioral changes etc. Hencethe material of the invention can be readily and safely administered tothe animals.

The route of administration of the material primarily depends on thesubject. Hence, if the subject is bovine species, then oral route may beadopted. Familiar methods of oral administration routes includesublingual, nasal, buccal. Other routes of administration, such ascutaneous, sub-cutaneous, parenteral, vaginal, intra-urethral, analroutes, etc. may also be adopted.

The material may be administered as such or may be formulated in variousphysical forms such as solution, syrup, elixir, mixture, emulsion,suspension, tablet, capsule, pessary, suppository, aerosol or a parentalpreparation, etc. The dosage form may accordingly be varied. As such,there is no intention to limit the scope of the invention to anyparticular physical form. In accordance with the practice of theinvention, pharmaceutical compositions containing the material as theprimary active ingredient may be prepared. These compositions may beprepared according to any method known in the art for the manufacture ofpharmaceutical compositions. Such compositions, if intended for oral usemay contain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents and preservingagents in order to provide pharmaceutically elegant and palatablepreparations. Tablets are prepared containing the active ingredient i.e.the material, in admixture with non-toxic pharmaceutically acceptableexcipients. Such excipients may be for example inert diluents, such ascalcium carbonate, sodium carbonate, lactose, calcium phosphate orsodium phosphate; granulating and disintegrating agents, for example,corn starch, or alginic acid; binding agents, such as starch, gelatin oracacia, and lubricating agents like magnesium stearate, stearic acid ortalc. The tablets may be uncoated or they may be coated by knowntechniques.

Compositions for oral use may also be presented as hard gelatin capsuleswherein the active ingredient i.e. the material is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient i.e.the material is mixed with water or an oil medium, for example peanutoil, liquid paraffin or olive oil.

The compositions may also be formulated as suppositories or pessarywhich can be prepared by mixing the material with suitable nonirritatingexcipients or carriers such as cocoa butter, polyethyleneglycol or asuppository wax, which are solid at ordinary temperatures but liquid atbody temperature and therefore, melt in the rectum or vaginal cavity andrelease the active ingredient i.e. the material. In other words, allsorts of compositions that do not affect the efficacy of the materialand are capable of keeping the active ingredient i.e. the material ineffective contact with the uterine tissues are envisaged and envisagedwithin the scope of this invention.

The exact mechanism by which the material i.e. vinegar, dilute aceticacid and their natural or synthetic, derivatives act is not clear.However, it can be postulated that these materials can furnish an acetylradical by certain biochemical interactions in the living system toparticipate in the Kreb's-cycle via the route of Acetyl-Co-Enzyme A. Buteven then the exact mechanism by which a change in normal metabolicpathway of the living system like Kreb's-cycle causes sex of theoffspring to be determined is not clear. And it may also happen thatabove material does not interfere in normal metabolic pathways of theliving systems but acts as a sex determinant by some other mechanism orbiochemical pathway, known or unknown, to present day availablescientific knowledge and data. Or, it may be that as certain reptilesand ratites do not have an organised sex chromatin and as they arephylogenic ancestors and relatives of mammals which are much evolvedchordates and as a prominent principle in evolution states that progenyrepeats phyllogeny; it may not be ruled out that under certaincircumstances and stages of fertilization or zygotic or embryonicdevelopment, a given chromosome, say X-chromosome or a chromatinpattern, say XX may change into the other type, i.e. Y-chromosome or XYchromatin pattern and vice versa in some sort of imitation to thephenomenon of birth reversal found in certain reptiles and ratites.

The foregoing description of the invention is considered illustrative ofsome of the preferred embodiments of the invention. Variousmodifications and changes that can be readily made by a person skilledin the art, are considered to be encompassed within the scope of thepresent invention. Accordingly, the embodiments illustrated above andthe following examples do not limit the scope of the invention to theexact features as herein described. Suitable modifications andequivalents may be resorted to, within the scope of the invention.

EXAMPLE 1 Preparation of Material

20 liters of sugarcane juice was obtained from crushed sugarcane. Thisjuice was left in an earthen pot in the open for fermentation for aperiod of 6 months. A fermenting agent such as vinegar preparedpreviously by the same process was added on day 1 and day 10 to thejuice in the earthen pot. The amount added was about 250 ml. Thetemperature prevalent during period was in the range of 25 to 35° C. Thejuice in the earthen pot was periodically monitored and decanted toensure that contaminants and unwanted organisms do not infect it. At theend of six months, 4 to 5 liters of liquid was found in the earthen pot.This liquid was tested for its contents. This liquid contained thefollowing:

Dilute Acetic acid 8 to 16% Acetaldehyde Traces Acetic anhydride TracesEthyl Alcohol 4 to 9%  Rest water to qs

EXAMPLE 2

The study was conducted as under:

-   -   (1) Study on animals (cows and buffaloes) and on control groups        using vinegar only.    -   (2) Study on animals (cows and buffaloes) with no control group        using vinegar, acetic acid and sodium acetate.    -   (3) Study on other animals using vinegar only.

The first study was conducted in a population primarily comprising cowsand buffaloes. The study had a total of 38 animals (26 buffaloes and 12cows). Equal number of animals were used in the standard/control groupfor comparison. The animals were in the study group were allowed toundergo one insemination. Thereafter, i.e. within 35-50 minutes, thematerial prepared according to the process described in Example 1 wasadministered to the animals in the study group. The animals in thecontrol/standard group were not subject to administration of anymaterial. Mating or insemination was not controlled. All the animalswere allowed to move freely in their usual environment and were kept onnormal diet. Their behavior and temperature/temperament especially ofthe study group. was monitored. It was found to be normal. No skinreaction, rashes, etc. were detected. No unnatural behavior was observedin any of the animals. All animals that conceived in the study and thecontrol group proceeded to pregnancy. Upon completion of pregnancy, itwas noted that out of 38 animals in the study group, 30 animalsdelivered female calves. During this period, it was also noted that 7did not conceive and one reported mis-carriage. In the control groupwhere no vinegar was administered, out of 38 animals (i.e. 26 buffaloesand 12 cows), 5 did not conceive and 1 reported miscarriage. Theremaining 32 animals delivered 16 males (46.87%) and 17 (53.13%) femaleoffsprings. The results are depicted in Table 1. It is to be noted thatthe cases of miscarriage and non-conception are not abnormal as it is ageneral phenomenon in these animals.

TABLE 1 Effects of vinegar on study and control groups of animalsAnimals Type of No. of No. of not OFFSPRING Animals Animals miscarriagesconceived Females Males STUDY Buffaloes 26 1 5 20 Nil Cows 12 Nil 2 10Nil STANDARD Buffaloes 26 1 3 12 10 Cows 12 Nil 2 5 5

EXAMPLE 3

A similar study, as described in Example 2, was carried out on animalssuch as cows and buffaloes using study groups. These animals wereadministered vinegar, dilute acetic acid, and sodium acetate solution inacidic pH separately. The results of this study are described hereinbelow and shown in Table 2.

-   A. Study using vinegar only: The study was carried on 21 animals (14    buffaloes and 7 cows) out of which 4 did not conceive and 1 reported    miscarriage and the remaining 16 delivered 16 (100%) female    offsprings.-   B. Study using Acetic acid only: The study was conducted on 22    animals (14 buffaloes and 8 cows) out of which 5 did not conceive;    no animals reported miscarriage and remaining the 17 animals    delivered 17 (100%) female offsprings.-   C. Study using sodium acetate solution in Acidic pH: The study was    conducted on 15 animals (10 buffaloes and 5 cows) out of which there    were 3 cases of no conception and remaining 12 animals delivered 12    (100%) female offsprings.

TABLE 2 Study on animals with no control group using vinegar acetic aciddilute and sodium acetate solution in acidic -pH Type of prepar- No. ofAnimals ation Type of No. of Miscar- not OFFSPRING used Animals Animalsriages conceived Females Males Vinegar Buffaloes 14 1 3 10 Nil Cows 7Nil 1 6 Nil Acetic Buffaloes 14 Nil 3 11 Nil Acid Cows 8 Nil 2 6 Nildilute Sodium Buffaloes 10 Nil 2 8 Nil Acetate Cows 5 Nil 1 4 Nil Acid-ified

EXAMPLE 4

In order to ascertain the efficacy of vinegar/acetic acid on othermammals, a study, as discussed in Examples 2 and 3 was conducted onsheep. The study had 20 sheep in the study group and 20 animals in thecontrol group. At the end of the study, it was found that the sheep inthe study group that were administered vinegar (50-60 ml/dose) gavebirth to 28 female offsprings. No male offspring was produced. 5 animalsdid not conceive and one animal reported miscarriage. The animals in thecontrol group, on the other hand, produced male and female offsprings.The results are shown in Table 3 herein below:

TABLE 3 Study on sheep using vinegar Animals not No. of OFFSPRING No. ofanimals conceived miscarriages Males Females Study 20 5 1 Nil 28 Control20 3 1 8 16

EXAMPLE 5

In order to ascertain the efficacy of vinegar/acetic acid on mammalssuch as horses, a study, as discussed in Examples 2 and 3 was conducted.The dosage of administration of vinegar was 250-400 ml. At the end ofthe study wherein 26 horses were employed, it was found that 17 femaleoffsprings were produced. No male offspring was produced. 7 animals didnot conceive and two animals reported miscarriage. The results are shownin Table 4 herein below:

TABLE 4 Study on horses using vinegar Animals not No. of OFFSPRING No.of animals conceived miscarriages Males Females 26 7 2 Nil 17

EXAMPLE 6

In order to ascertain the efficacy of vinegar/acetic acid on goats, astudy, as discussed in Examples 2 and 3 was conducted. The study had 19goats. The animals were given 50-60 ml of vinegar after theinsemination. At the end, of the study wherein 19 goats were employed,it was found that 26 female offsprings were produced. No male offspringwas produced by the goats. 6 animals did not conceive and no cases ofmiscarriage were reported. The results are shown in Table 5, hereinbelow:

TABLE 5 Study on goats using vinegar Animals not No. of OFFSPRING No. ofanimals conceived miscarriages Males Females 19 6 Nil Nil 26

EXAMPLE 7

Yet another study on pigs was conducted following the method discussedin Example 2 and 3. The pigs were given 150 to 250 ml of vinegar. At theend of the study wherein 7 pigs were employed, it was found that 47female offsprings were produced. No male offspring was produced. 2animals did not conceive and no cases of miscarriage were reported. Theresults are shown in Table 6, herein below:

TABLE 6 Study on pigs using vinegar only Animals not No. of OFFSPRINGNo. of animals conceived miscarriages Males Females 7 2 Nil Nil 47

EXAMPLE 8

A study on dogs based on the method described in Examples 2 and 3 wasconducted on dogs. Vinegar was given to all the animals @45-55 ml. Atthe end of the study wherein 9 dogs were employed, it was found that 29female offsprings were produced. No male offspring was produced. 2animals did not conceive and no cases of miscarriage were reported. Theresults are shown in Table 7, herein below:

TABLE 7 Study on dogs using vinegar Animals not No. of OFFSPRING No. ofanimals conceived miscarriages Males Females 9 2 Nil Nil 29

EXAMPLE 9

7 persons who had no female children in their family, came forth asvolunteers. They were informed about the study conducted in animals andthe efficacy of the material. The method, mode and dosage ofadministration (65-80 ml) was also discussed. The volunteers co-operatedand at the end of the study, 7 girl children were born. No males orcases of miscarriage were reported. The results are shown in Table 8,herein below:

TABLE 8 Study on humans using vinegar No. of human No. of OFFSPRINGsubjects miscarriages Males Females 7 Nil Nil 7

1. An in vivo method for the preferential production of female offspringin a bovine, said method comprising the step of orally administering amaterial comprising a therapeutically effective amount of acetic acidand/or its pharmaceutically acceptable derivatives to the bovine in atime period defined as 3 hours before insemination of the bovine to 1hour after insemination of the bovine, provided that the material isadministered to the bovine before zygote formation.
 2. The method asclaimed in claim 1 wherein the material comprises vinegar, sodiumacetate and/or their natural or synthetic derivatives.
 3. The method asclaimed in claim 1 wherein the material comprises 8-16% dilute aceticacid.
 4. The method as claimed in claim 1 wherein the amount of thematerial administered to the bovine is in the range of 150 ml to 800 ml.5. The method as claimed in claim 1 wherein the amount of the materialadministered to the bovine is in the range of 250 ml to 400 ml.
 6. Themethod as claimed in claim 1 wherein the material is administered afterinsemination but before zygote formation.
 7. The method as claimed inclaim 1 wherein the material is formulated in a physical form selectedfrom the group consisting of a solution, a syrup, an elixir, a mixture,an emulsion, a suspension, a tablet, and a capsule.
 8. The method asclaimed in claim 1 wherein the material is administered within threehours before zygote formation.
 9. The method as claimed in claim 1wherein the material is administered to the bovine within 30 minutesafter insemination of the bovine.
 10. The method as claimed in claim 1wherein the material is administered to the bovine one hour beforeinsemination of the bovine.